title: Mouse cloning using a drop of peripheral blood.
abstract: Somatic cell nuclear transfer (SCNT) is a unique technology that produces cloned animals from single cells. It is desirable from a practical viewpoint that donor cells can be collected noninvasively and used readily for nuclear transfer. The present study was undertaken to determine whether peripheral blood cells freshly collected from living mice could be used for SCNT. We collected a drop of peripheral blood (15-45 μl) from the tail of a donor. A nucleated cell (leukocyte) suspension was prepared by lysing the red blood cells. Following SCNT using randomly selected leukocyte nuclei, cloned offspring were born at a 2.8% birth rate. Fluorescence-activated cell sorting revealed that granulocytes/monocytes and lymphocytes could be roughly distinguished by their sizes, the former being significantly larger. We then cloned putative granulocytes/monocytes and lymphocytes separately and obtained 2.1% and 1.7% birth rates, respectively (P > 0.05). Because the use of lymphocyte nuclei inevitably results in the birth of offspring with DNA rearrangements, we applied granulocyte/monocyte cloning to two genetically modified strains and two recombinant inbred strains. Normal-looking offspring were obtained from all four strains tested. The present study clearly indicated that genetic copies of mice could be produced using a drop of peripheral blood from living donors. This strategy will be applied to the rescue of infertile founder animals or a "last-of-line" animal possessing invaluable genetic resources.
authors.affiliation: RIKEN BioResource Center, Tsukuba, Ibaraki, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kamimura
authors.forename: Kimiko
authors.initials: K
authors.lastname: Inoue
authors.forename: Narumi
authors.initials: N
authors.lastname: Ogonuki
authors.forename: Michiko
authors.initials: M
authors.lastname: Hirose
authors.forename: Mami
authors.initials: M
authors.lastname: Oikawa
authors.forename: Masahiro
authors.initials: M
authors.lastname: Yo
authors.forename: Osamu
authors.initials: O
authors.lastname: Ohara
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Miyoshi
authors.forename: Atsuo
authors.initials: A
authors.lastname: Ogura
mesh_terms: D000818:Animals; D019976:Cloning, Organism / Q000379:methods*; D004624:Embryo Transfer; D004622:Embryo, Mammalian / Q000166:cytology*; D005347:Fibroblasts / Q000166:cytology; D005434:Flow Cytometry; D051379:Mice; D053652:Nuclear Transfer Techniques
keywords: granulocyte; leukocyte; lymphocyte; mouse; somatic cell nuclear transfer